Comparison of three methods for virus enrichment from purified water

Abstract

ObjectiveTo compare the effects and experimental conditions of membrane adsorption method， membrane adsorption-PEG method and membrane adsorption-ultrafiltration method on virus enrichment in purified water.MethodsMS2 bacteriophage was used as target virus to prepare water samples with high and low concentrations， and the standard curve of reverse transcription-fluorescence quantitative polymerase chain reaction （RT-qPCR） was established for quantitative detection. Water samples were enriched by membrane adsorption method， membrane adsorption-PEG method and membrane adsorption-ultrafiltration method. The recoveries of different filtration membranes， different ultrafiltration tubes and two nucleic acid extraction methods were compared.ResultsThe average recoveries of membrane adsorption-ultrafiltration method， membrane adsorption-PEG method and membrane adsorption method were 19.80% ± 12.19%， 12.71% ± 9.09% and 9.05% ± 4.89%. The order of influencing factors on the recoveries was： membrane adsorption-ultrafiltration method > membrane adsorption-PEG method > membrane adsorption method. High concentration > low concentration. The effect of virus enrichment of mixed cellulose ester membrane was better than nylon membrane. There was no difference in the enrichment effect of ultrafiltration tube at 100 and 50 K， and the nucleic acid extraction effect of silica gel centrifugation column method and magnetic bead method.ConclusionThe enrichment efficiency can be improved by secondary enrichment. Membrane adsorption-ultrafiltration method is simple to operate， short time consuming， low dependence on equipment， and easy to be popularized. The experimental conditions of membrane adsorption-ultrafiltration can be optimized and improved to expand the application of different water bodies and different virus enrichment.

Keywords

• waterborne virus
• enrichment
• membrane adsorption
• ultrafiltration